title: Inhibition of chemotactic motility and trans-endothelial migration of human neutrophils by sphingosine 1-phosphate.
abstract: In previous studies, we reported that sphingosine 1-phosphate (Sph-1-P) inhibits the chemotactic motility of some cancer cell lines such as mouse melanoma cells, as well as human smooth muscle cells, at a very low concentration, as demonstrated by a transwell migration assay method (Proc. Natl. Acad. Sci. USA 89, 9698, 1992; J. Cell Biol. 130, 193, 1995). In this study, we investigated the effect of Sph-1-P on the chemotactic motility and invasiveness of human neutrophils, utilizing three different assay systems: (a) a transwell migration assay where IL-8 or fLMP was added as a chemotactic factor, (b) a phagokinetic assay with gold colloids, and (c) a trans-endothelial migration assay with human umbilical vein endothelial cells (HUVECs) plated on collagen layers. We found that among various sphingosine derivatives, Sph-1-P specifically inhibited the IL-8- or fLMP-induced chemotactic migration of neutrophils at concentrations below 1 microM. Phagokinetic activity of neutrophils was also suppressed by Sph-1-P, but more moderately than by the PKC inhibitory sphingosine analog, trimethylsphingosine. Finally, Sph-1-P inhibited trans-endothelial migration and invasiveness of neutrophils into HUVEC-covered collagen layers, whereas no effect on their adhesion to HUVECs was observed. These observations strongly suggest that Sph-1-P can act as a specific and effective motility regulator of human neutrophils, raising the possibility of future applications of Sph-1-P, or its analogs, as anti-inflammatory agents regulating invasive migration of neutrophils through endothelial layers at injured vascular sites.
authors.affiliation: The Biomembrane Institute and Department of Pathobiology, University of Washington, Seattle 90104, USA.
authors.forename: S
authors.initials: S
authors.lastname: Kawa
authors.forename: S
authors.initials: S
authors.lastname: Kimura
authors.forename: S
authors.initials: S
authors.lastname: Hakomori
authors.forename: Y
authors.initials: Y
authors.lastname: Igarashi
mesh_terms: D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects*; D002478:Cells, Cultured; D002518:Ceramides / Q000494:pharmacology; D002633:Chemotaxis / Q000187:drug effects*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D017979:Gold Colloid; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D016209:Interleukin-8 / Q000494:pharmacology; D008246:Lysophospholipids; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils; D010587:Phagocytosis; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D014470:Umbilical Cord
chemical_list: D002518:Ceramides; D004791:Enzyme Inhibitors; D017979:Gold Colloid; D016209:Interleukin-8; D008246:Lysophospholipids; C071600:N,N,N-trimethylsphingosine; C060506:sphingosine 1-phosphate; D009240:N-Formylmethionine Leucyl-Phenylalanine; D011493:Protein Kinase C; D013110:Sphingosine